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Updated: Jun 21, 2025
12:04
Mouse Genome Engineering Using Designer Nucleases
Published on: April 2, 2014
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Large-Scale Genome-Wide Optimization and Prediction of the Cre Recombinase System for Precise Genome Manipulation in
Vishnu Hosur1, Valerie Erhardt1, Elli Hartig1
1The Jackson Laboratory for Mammalian Genetics.
Research Square
|July 16, 2024
Summary
Optimizing Cre-Lox recombination in mice requires careful consideration of factors like loxP site distance and Cre-driver strain selection. Understanding these elements improves efficiency and reduces costs in generating genetically modified mouse models.
Area of Science:
- Genetics and Genomics
- Molecular Biology
- Animal Models
Background:
- The Cre-Lox recombination system is vital for conditional gene manipulation in mouse genetics.
- Challenges include high costs, time investment, and unpredictable recombination events.
- Systematic analysis is needed to optimize Cre-Lox efficiency for generating mouse models.
Purpose of the Study:
- To conduct a genome-wide analysis of factors influencing Cre-mediated recombination in mice.
- To identify optimal conditions for efficient and reproducible Cre-Lox recombination.
- To provide guidance for generating conditional knockout mice and other genetic models.
Main Methods:
- Generated 11 novel mouse strains with conditional alleles at the ROSA26 locus on a C57BL/6J background.
- Systematically evaluated the impact of inter-loxP distance, mutant loxP sites, zygosity, chromosomal location, and breeder age on recombination.
- Utilized Cre-driver strains to assess recombination efficiency under various conditions.
Main Results:
- Cre-driver strain selection significantly impacts recombination efficiency.
- Optimal recombination occurs with loxP sites 1-4 kb apart; mutant loxP sites are effective at 1-3 kb.
- Complete recombination is hindered at distances ≥15 kb (wildtype loxP) or ≥7 kb (mutant loxP); breeder age (8-20 weeks) and heterozygous alleles improve efficiency.
Conclusions:
- Inter-loxP distance, loxP site type, zygosity, and breeder age are critical for efficient Cre-Lox recombination.
- Maintaining an inter-loxP distance ≤4 kb ensures efficient recombination regardless of chromosomal location.
- This study provides essential data to optimize Cre-Lox strategies, reducing time and cost in generating mouse models.
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